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An economic, square-shaped flat-field illumination module for TIRF-based super-resolution microscopy.

ABSTRACT: Super-resolution microscopy allows complex biological assemblies to be observed with remarkable resolution. However, the presence of uneven Gaussian-shaped illumination hinders its use in quantitative imaging or high-throughput assays. Methods developed to circumvent this problem are often expensive, hard to implement, or not applicable to total internal reflection fluorescence imaging. We herein demonstrate a cost-effective method to overcome these challenges using a small square-core multimodal optical fiber as the coupler. We characterize our method with synthetic, recombinant, and cellular systems imaged under total internal reflection fluorescence and highly inclined and laminated optical sheet illuminations to demonstrate its ability to produce highly uniform images under all conditions.

SUBMITTER: Lam JYL

PROVIDER: S-EPMC8914601 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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An economic, square-shaped flat-field illumination module for TIRF-based super-resolution microscopy.

Lam Jeff Y L JYL Wu Yunzhao Y Dimou Eleni E Zhang Ziwei Z Cheetham Matthew R MR Körbel Markus M Xia Zengjie Z Klenerman David D Danial John S H JSH

Biophysical reports 20220309 1

Super-resolution microscopy allows complex biological assemblies to be observed with remarkable resolution. However, the presence of uneven Gaussian-shaped illumination hinders its use in quantitative imaging or high-throughput assays. Methods developed to circumvent this problem are often expensive, hard to implement, or not applicable to total internal reflection fluorescence imaging. We herein demonstrate a cost-effective method to overcome these challenges using a small square-core multimoda ...[more]

PMID: 35299715

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